U2OS Cells GFP-NUP98
SIGMA/CLL1136
Product Type: Chemical
Fluorescence Microscopy U2OS GFP-NUP98: Cat. No. CLL1136: U2OS osteosarcoma knock-in cell line expressing the NUP98 gene (nucleoporin 98 kDa, a peripheral component of the NPC) endogenously tagged with 3xFLAG-GFP at the N-terminus. Cells were imaged live in Hanks Balanced Salt solution, Cat. No. H8264, supplemented with 2% Fetal Bovine Serum, Cat. No. F2442, using GFP filterset (ex 450–490/em 500–550) and 40×/1.3 oil objective.
Differential Interference Contrast (DIC) Microscopy U2OS GFP-NUP98: Cat. No. CLL1136: Differential interference contrast (DIC) microscopy of the same cells seen in the fluorescence microscopy image.
Fluorescence Microscopy U2OS GFP-NUP98: Cat. No. CLL1136: Another fluorescence image of the same cells seen in the fluorescence microscopy and DIC images. In this image the focal plane was moved down so that it touches the lower surface of the nuclei near the glass. This is in contrast to the main fluorescence image, in which the focal plane crosses the middle of the nuclei.
| biological source | human female bone (Source Disease: Bone osteosarcoma) |
| OMIM accession no. | 601021   |
| Quality Level | 200 |
| storage temp. | −196°C |
| Analysis Note: | Tested for Mycoplasma, sterility, post-freeze viability, short terminal repeat (STR) analysis for cell line identification, cytochrome oxidase I (COI) analysis for cell line species confirmation. |
| Application: | U2OS Cells have been used to assess cell proliferation using Alamar Blue and live cell imaging. This product is a human U2OS cell line in which the genomic NUP98 gene has been endogenously tagged with a 3XFLAG™ epitope and a Green Fluorescent Protein (GFP) gene using CompoZr® Zinc Finger Nuclease technology. Integration resulted in endogenous expression of a fusion protein in which the 3XFLAG is attached to the N-terminus of GFP and the GFP is attached to the N-terminus of NUP98. Fluorescence imaging shows characteristic NUP98 nuclear expression. This stable cell line was expanded from a single clone. The target’s gene regulation and corresponding protein function are preserved in contrast to cell lines with overexpression via an exogenous promoter. To learn more, please visit the Cellular Reporter Cell Line webpage |
| Disclaimer: | RESEARCH USE ONLY. This product is regulated in France when intended to be used for scientific purposes, including for import and export activities (Article L 1211-1 paragraph 2 of the Public Health Code). The purchaser (i.e. enduser) is required to obtain an import authorization from the France Ministry of Research referred in the Article L1245-5-1 II. of Public Health Code. By ordering this product, you are confirming that you have obtained the proper import authorization. |
| Features and Benefits: | Zinc finger nuclease (ZFN)-mediated targeted integration of a fluorescent GFP tag just behind the start codon of the NUP98 gene on chromosome 11p15.4 to create a cell line exhibiting stable expression of the transgene tagged with GFP on the N-terminus of the protein. The U2OS cells are adherent, with a doubling time of approx. 29 hours. |
| General description: | U2OS GFP-NUP98 are bone osteosarcoma cells from a human 15 year old caucasian female having a ZFN modification creating a GFP-NUP98 transgene expressed from the endogenous NUP98 gene locus. This cell line was derived from ATCC Catalog No. HTB-96. |
| Legal Information: | 3xFLAG is a trademark of Sigma-Aldrich Co. LLC |
| Legal Information: | CompoZr is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Preparation Note: | Media Renewal changes two to three times per week. Rapidly thaw vial by gentle agitation in 37°C water bath (~2 minutes), keeping vial cap out of the water. Decontaminate with 70% ethanol, add 9 mL culture media and centrifuge 125 x g (5-7 minutes). Resuspend in complete culture media and incubate at 37°C in a 5% CO2 atmosphere. Subculture Ratio: approx. 1:3-1:6 The base medium for this cell line is McCoy′s 5A Medium Modified, Cat. No. M9309. To make the complete growth medium, add the following components to the base medium: fetal bovine serum, Cat. No. F4135, to a final concentration (v/v) of 10%. Cell freezing medium-DMSO 1X, Cat. No. C6164. |
| WGK Germany | WGK 3 |
| Storage Temp. | −196°C |
| UNSPSC | 41106514 |